tIn this study we aimed to identify genes that are responsive to pertussis toxin (PTx) and might eventu-ally be used as biological markers in a testing strategy to detect residual PTx in vaccines. By microarrayanalysis we screened six human cell types (bronchial epithelial cell line BEAS-2B, fetal lung fibroblastcell line MRC-5, primary cardiac microvascular endothelial cells, primary pulmonary artery smooth mus-cle cells, hybrid cell line EA.Hy926 of umbilical vein endothelial cells and epithelial cell line A549 andimmature monocyte-derived dendritic cells) for differential gene expression induced by PTx. Imma-ture monocyte-derived dendritic cells (iMoDCs) were the only cells in which PTx induced significantdifferential expression of genes. Results were confirmed using different donors and further extendedby showing specificity for PTx in comparison to Escherichia coli lipopolysaccharide (LPS) and Bordetellapertussis lipo-oligosaccharide (LOS). Statistical analysis indicated 6 genes, namely IFNG, IL2, XCL1, CD69,CSF2 and CXCL10, as significantly upregulated by PTx which was also demonstrated at the protein levelfor genes encoding secreted proteins. IL-2 and IFN- gave the strongest response. The minimal PTx con-centrations that induced production of IL-2 and IFN- in iMoDCs were 12.5 and 25 IU/ml, respectively.High concentrations of LPS slightly induced IFN- but not IL-2, while LOS and detoxified pertussis toxindid not induce production of either cytokine. In conclusion, using microarray analysis we evaluated sixhuman cell lines/types for their responsiveness to PTx and found 6 PTx-responsive genes in iMoDCs ofwhich IL2 is the most promising candidate to be used as a biomarker for the detection of residual PTx.
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Abstract from the article: "Conjugates of fatty acids and amines, including endocannabinoids, are known to play important roles as endogenous signaling molecules. Among these, the ethanolamine conjugate of the n-3 poly unsaturated long chain fatty acid (PUFA) docosahexaenoic acid (22:6n-3) (DHA) was shown to possess strong anti-inflammatory properties. Previously, we identified the serotonin conjugate of DHA, docosahexaenoyl serotonin (DHA-5-HT), in intestinal tissues and showed that its levels are markedly influenced by intake of n-3 PUFAs. However, its biological roles remain to be elucidated. Here, we show that DHA-5-HT possesses potent anti-inflammatory properties by attenuating the IL-23-IL-17 signaling cascade in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Transcriptome analysis revealed that DHA-5-HT down-regulates LPS-induced genes, particularly those involved in generating a CD4+ Th17 response. Hence, levels of PGE2, IL-6, IL-1β, and IL-23, all pivotal macrophage-produced mediators driving the activation of pathogenic Th17 cells in a concerted way, were found to be significantly suppressed by concentrations as low as 100–500 nM DHA-5-HT. Furthermore, DHA-5-HT inhibited the ability of RAW264.7 cells to migrate and downregulated chemokines like MCP-1, CCL-20, and gene-expression of CCL-22 and of several metalloproteinases. Gene set enrichment analysis (GSEA) suggested negative overlap with gene sets linked to inflammatory bowel disease (IBD) and positive overlap with gene sets related to the Nrf2 pathway. The specific formation of DHA-5-HT in the gut, combined with increasing data underlining the importance of the IL-23-IL-17 signaling pathway in the etiology of many chronic inflammatory diseases merits further investigation into its potential as therapeutic compound in e.g. IBD or intestinal tumorigenesis".
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Background: Chronic low-grade inflammatory profile (CLIP) is one of the pathways involved in type 2 diabetes (T2D). Currently, there is limited evidence for ameliorating effects of combined lifestyle interventions on CLIP in type 2 diabetes. We investigated whether a 13-week combined lifestyle intervention, using hypocaloric diet and resistance exercise plus high-intensity interval training with or without consumption of a protein drink, affected CLIP in older adults with T2D. Methods: In this post-hoc analysis of the PROBE study 114 adults (≥55 years) with obesity and type 2 (pre-)diabetes had measurements of C-reactive protein (CRP), pro-inflammatory cytokines interleukin (IL)-6, tumor-necrosis-factor (TNF)-α, and monocyte chemoattractant protein (MCP)-1, anti-inflammatory cytokines IL-10, IL-1 receptor antagonist (RA), and soluble tumor-necrosis-factor receptor (sTNFR)1, adipokines leptin and adiponectin, and glycation biomarkers carboxymethyl-lysine (CML) and soluble receptor for advanced glycation end products (sRAGE) from fasting blood samples. A linear mixed model was used to evaluate change in inflammatory biomarkers after lifestyle intervention and effect of the protein drink. Linear regression analysis was performed with parameters of body composition (by dual-energy X-ray absorptiometry) and parameters of insulin resistance (by oral glucose tolerance test). Results: There were no significant differences in CLIP responses between the protein and the control groups. For all participants combined, IL-1RA, leptin and adiponectin decreased after 13 weeks (p = 0.002, p < 0.001 and p < 0.001), while ratios TNF-α/IL-10 and TNF-α/IL-1RA increased (p = 0.003 and p = 0.035). CRP increased by 12 % in participants with low to average CLIP (pre 1.91 ± 0.39 mg/L, post 2.13 ± 1.16 mg/L, p = 0.006) and decreased by 36 % in those with high CLIP (pre 5.14 mg/L ± 1.20, post 3.30 ± 2.29 mg/L, p < 0.001). Change in leptin and IL-1RA was positively associated with change in fat mass (β = 0.133, p < 0.001; β = 0.017, p < 0.001) and insulin resistance (β = 0.095, p = 0.024; β = 0.020, p = 0.001). Change in lean mass was not associated with any of the biomarkers. Conclusion: 13 weeks of combined lifestyle intervention, either with or without protein drink, reduced circulating adipokines and anti-inflammatory cytokine IL-1RA, and increased inflammatory ratios TNF-α/IL-10 and TNF-α/IL-1RA in older adults with obesity and T2D. Effect on CLIP was inversely related to baseline inflammatory status.
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Background and aims: Observational data indicate that diets rich in fruits and vegetables have a positive effect on inflammatory status, improve metabolic resilience and may protect against the development of non-communicable diseases. Nevertheless, experimental evidence demonstrating a causal relationship between nutrient intake (especially whole foods) and changes in metabolic health is scarce. This study investigated the pleiotropic effects of sulforaphane from broccoli sprouts, compared to pea sprouts, on biomarkers of endothelial function, inflammation and metabolic stress in healthy participants subjected to a standardized caloric challenge.Methods: In this double-blind, crossover, randomized, placebo-controlled trial 12 healthy participants were administered 16 g broccoli sprouts, or pea sprouts (placebo) followed by the standardized high-caloric drink PhenFlex given to disturb healthy homeostasis. Levels of inflammatory biomarkers and metabolic parameters were measured in plasma before and 2 h after the caloric overload.Results: Administration of broccoli sprouts promoted an increase in levels of CCL-2 induced by caloric load (p = 0.017). Other biomarkers (sICAM-1, sVCAM-1, hs-CRP, and IL-10) individually showed insignificant tendencies toward increase with administration of sulforaphane. Combining all studied biomarkers into the systemic low-grade inflammation score further confirmed upregulation of the inflammatory activity (p = 0.087) after sulforaphane. No significant effects on biomarkers of metabolic stress were detected.Conclusion: This study has demonstrated that sulforaphane facilitated development of a mild pro-inflammatory state during the caloric challenge, which could be suggestive of the onset of the hormetic response induced by this phytonutrient. The use of integrative outcomes measures such as the systemic low-grade inflammation score can be viewed as a more robust approach to study the subtle and pleiotropic effects of phytonutrients.Clinical trial registration:www.clinicaltrials.gov, identifier NCT05146804.Keywords: biomarkers; diet; glucoraphanin; hormesis; inflammation; nutrients; phenotypic flexibility; sulforaphane.
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BACKGROUND: T lymphocytes are orchestrators of adaptive immunity. Naïve T cells may differentiate into Th1, Th2, Th17 or iTreg phenotypes, depending on environmental co-stimulatory signals. To identify genes and pathways involved in differentiation of Jurkat T cells towards Th1 and Th2 subtypes we performed comprehensive transcriptome analyses of Jurkat T cells stimulated with various stimuli and pathway inhibitors. Results from these experiments were validated in a human experimental setting using whole blood and purified CD4+ Tcells.RESULTS: Calcium-dependent activation of T cells using CD3/CD28 and PMA/CD3 stimulation induced a Th1 expression profile reflected by increased expression of T-bet, RUNX3, IL-2, and IFNγ, whereas calcium-independent activation via PMA/CD28 induced a Th2 expression profile which included GATA3, RXRA, CCL1 and Itk. Knock down with siRNA and gene expression profiling in the presence of selective kinase inhibitors showed that proximal kinases Lck and PKCθ are crucial signaling hubs during T helper cell activation, revealing a clear role for Lck in Th1 development and for PKCθ in both Th1 and Th2 development. Medial signaling via MAPkinases appeared to be less important in these pathways, since specific inhibitors of these kinases displayed a minor effect on gene expression. Translation towards a primary, whole blood setting and purified human CD4+ T cells revealed that PMA/CD3 stimulation induced a more pronounced Th1 specific, Lck and PKCθ dependent IFNγ production, whereas PMA/CD28 induced Th2 specific IL-5 and IL-13 production, independent of Lck activation. PMA/CD3-mediated skewing towards a Th1 phenotype was also reflected in mRNA expression of the master transcription factor Tbet, whereas PMA/CD28-mediated stimulation enhanced GATA3 mRNA expression in primary human CD4+ Tcells.CONCLUSIONS: This study identifies stimulatory pathways and gene expression profiles for in vitro skewing of T helper cell activation. PMA/CD3 stimulation enhances a Th1-like response in an Lck and PKCθ dependent fashion, whereas PMA/CD28 stimulation results in a Th2-like phenotype independent of the proximal TCR-tyrosine kinase Lck. This approach offers a robust and fast translational in vitro system for skewed T helper cell responses in Jurkat T cells, primary human CD4+ Tcells and in a more complex matrix such as human whole blood.
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Achtergrond: Ernstige psychische aandoeningen (epa) gaan gepaard met hoge ggz- en andere zorgkosten. Om de discussie hierover tussen verzekeraars, gemeenten en ggz transparanter te voeren zijn in 2012 zorgvignetten ontwikkeld die in een regio een beeld moeten geven over de aanwezige groepen patiënten met epa, ingedeeld naar zorgintensiteit. Doel: Inzicht krijgen in de vraag hoe bruikbaar en valide deze epa-vignetten zijn in relatie tot zorgbehoeften en psychosociaal functioneren. Methode: rom-uitkomsten (zorgbehoeften en psychosociaal functioneren) van 706 patiënten werden per zorgzwaartevignet vergeleken (m.b.v. χ2-toetsen en anova’s). Voor twee complexe zorgzwaartevignetten (zorgwekkende zorgmijders en gevaar afwenden) werden verschillen in zorgbehoeften en functioneren longitudinaal onderzocht (met repeated measures-anova’s en analyses volgens McNemar). Resultaten: De vignetten waren in beperkte mate onderscheidend wat betreft zorgbehoeften en functioneren. Patiënten met vignetten ‘zorgwekkende zorgmijders’ en ‘gevaar afwenden’ hadden de meeste beperkingen in functioneren en de meeste (onvervulde) zorgbehoeften. Deze bevindingen bleken stabiel over tijd. Conclusie: De vignetten zijn onvoldoende onderscheidend. Om regionale zorgplanning te verbeteren is het wenselijk om bestaande informatie van zorgvignetten te verrijken met informatie over zorgbehoeften en functioneren
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De LO-docent is vaak niet de enige beweegprofessional in en rondom een school die zich bezighoudt met beweegstimulering van kinderen om ze voldoende te laten bewegen. Ook bijvoorbeeld een buurtsportcoach of JOGG-regisseur kunnen hier bij betrokken zijn. Iedere professional heeft zijn eigen invalshoek van waaruit hij/zij werkt met bepaalde taken en verantwoordelijkheden. Die taken zijn niet alleen afhankelijk van welke beweegprofessionals allemaal betrokken zijn, maar ook van de unieke context van de school met daarin de school-specifieke behoeften, wensen en mogelijkheden. Om inzicht te krijgen in het veelzijdige werk van de beweegprofessionals in en rondom school hebben we zes rollen ontwikkeld, gebaseerd op de eerder ontwikkelde rollen van de Gezonde School-adviseur. In dit artikel beschrijven we deze zes rollen en gaan we vervolgens in op de mogelijkheden die deze rollen bieden voor zowel de verdere professionalisering van de beweegprofessional, als voor de samenwerking op schoolniveau.
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Le Premier ministre néerlandais Mark Rutte annonce en septembre 2014 la fin de l’État-providence et l’avènement de la société participative. Dans cette nouvelle société, chaque citoyen participe activement à la vie sociale et économique. Les citoyens y prennent par exemple la responsabilité de l’entretien de la rue et des espaces verts du quartier. Ils s’entraident et offrent du soutien et des soins aux voisins âgés, malades ou handicapés. Chaque individu y tient sa place dans la mesure de ses capacités personnelles. Par cette démarche, l’aide sociale n’est pas d’abord l’affaire de professionnels et de services sociaux, mais celle d’un réseau informel de personnes présentes dans l’entourage de la personne concernée. En effet, chaque citoyen, patient ou usager est invité à prendre en charge sa propre vie, pour le bien comme pour le mal. Il s’agit d’une dynamique de « Do-it-yourself-care », « le-social-fait-par-vous-même »1. Or, cette attitude du « Do-it-yourself » est aussi revendiquée par les entrepreneurs sociaux qui s’engagent par le bas dans les quartiers selon les principes de la nouvelle économie du partage. Dans ce chapitre, on se concentrera sur la figure de l’entrepreneur qui joue un rôle clé dans le renversement du système social néerlandais en cours. On y voit apparaître la logique du sujet2 déclinée dans la dynamique de subjectivation et de désubjectivation telle qu’évoquée par Michel Wieviorka3.
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In Nederland is er niet alleen een lerarentekort, er is ook een tekort aan masteropgeleide leraren. In de LO misschien nog wel meer dan bij andere vakken. Maar waarom zou je als leraar LO een master gaan doen, je hebt toch al een eerstegraadsbevoegdheid? |
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Materiaal ontwikkeld in het kader van het promotieonderzoek van Charline Rouffet naar de invloed van communicatieve toetsing op het vakdidactisch handelen van docenten moderne vreemde talen.
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